Large‐scale separation of acetylcholinesterase inhibitors from Zanthoxylum nitidum by pH‐zone‐refining counter‐current chromatography target‐guided by ultrafiltration high‐performance liquid chromatography with ultraviolet and mass spectrometry screening

A new strategy by converging ultrafiltration high‐performance liquid chromatography with ultraviolet and mass spectrometry and pH‐zone‐refining counter‐current chromatography was developed for the rapid screening and separation of potential acetylcholinesterase inhibitors from the crude alkaloidals extract of Zanthoxylum nitidum. An optimized two‐phase solvent system composed of chloroform/methanol/water (4:3:3, v/v) was used in this study. And, in the optimal solvent system, 45 mM hydrochloric acid was added to the aqueous stationary phase as the retainer, while 5 mM triethylamine was added to the organic mobile phase as the eluter. As a result, with the purity of over 95%, five alkaloids including jatrorrhizine ( 1 , 340 mg), columbamine ( 2 , 112 mg), skimmianine ( 3 , 154 mg), palmatine ( 4 , 226 mg), and epiberberine ( 5 , 132 mg) were successfully purified in one step from 3.0 g crude alkaloidals extract. And their structures were identified by ultraviolet, mass spectrometry, ¹H and ¹³C NMR spectroscopy. Notably, compounds 2 , 4 and 5 were firstly reported in Z. nitidum. In addition, acetylcholinesterase inhibitory activities of compounds 1–5 were evaluated, and compounds 3, 4 and 5 exhibited stronger acetylcholinesterase inhibitory activity (IC₅₀ values at 8.52 ± 0.64, 14.82 ± 1.21 and 3.12 ± 0.32 μg/mL, respectively) than berberine (IC₅₀ value at 32.86 ± 2.14 μg/mL, positive control). The results indicated that the proposed method is an efficient technique to rapidly screen acetylcholinesterase inhibitors from complex samples, and could be served as a large‐scale preparative technique for separating ionizable active compounds.     

GC-MS analysis of metabolites from soxhlet extraction,ultrasound-assisted extraction and supercritical fluid extraction of Salacca zalacca flesh and its alpha-glucosidase inhibitory activity

Abstract

Different extraction processes were employed to extract bioactive metabolites from Salacca zalacca flesh by a range of aqueous and organic solvents. The highest extraction yield was obtained by 50% ethanol extract of SE (73.18?±?4.35%), whereas SFE_1 showed the lowest yield (0.42?±?0.08%). All extracts were evaluated for in vitro α-glucosidase inhibitory activity, measured by their IC₅₀ values in comparison to that of quercetin, the positive control (IC₅₀ = 2.7?±?0.7?μg/mL). The lowest α-glucosidase inhibitory activity was indicated by water extract of SE (IC₅₀ = 724.3?±?42.9?μg/mL) and the highest activity was demonstrated by 60% ethanol extract by UAE (IC₅₀ = 16.2?±?2.4?μg/mL). All extracts were analysed by GC-MS and identified metabolites like carbohydrates, fatty acids, organic acids, phenolic acids, sterols and alkane-based compounds etcetera that may possess the potential as α-glucosidase inhibitor and may attribute to the α-glucosidase inhibitory activity.     

Chemical Constituents from the Stems of Mahonia Japonica

A new dihydroberberine alkaloid, 7,8‐dihydro‐8‐methoxyberberine ( 1 ), along with six known compounds including two dihydroberberine alkaloids, 7,8‐dihydro‐8‐hydroxyberberine ( 2 ) and oxyberberine ( 3 ) and four protoberberine alkaloids, berberine ( 4 ), palmatine ( 5 ), jatrorrhizine ( 6 ) and columbamine ( 7 ), were isolated from the stems of Mahonia japonica. These compounds were characterized and identified by physical and spectral evidence.     

Identification and analysis of alkaloids in cortex Phellodendron amurense by high‐performance liquid chromatography with electrospray ionization mass spectrometry coupled with photodiode array detection

Alkaloids from Cortex Phellodendron amurense Rupr. were identified to determine the material basis for the bioactivity of this herb. HPLC–ESI‐MS with photodiode array detection coupled to XCharge C₁₈ column was applied to analyze the alkaloids qualitatively and quantitatively. A total of 37 alkaloids were identified and tentatively characterized from the ethanol extract by online ESI‐MSⁿ fragmentation and UV spectral analysis. A total of ten alkaloids, including four novel natural products, were tentatively identified for the first time in P. amurense. The fragmentation pathways for certain compounds were analyzed. The contents of a pair of isomers (columbamine and jatrorrhizine) and four main alkaloids (phellodendrine, magnoflorine, berberine, and palmatine) were simultaneously quantified using the aforementioned method. Results showed that the newly discovered and known components of P. amurense were helpful in determining the material basis for the bioactivity of the herb. The application of the XCharge C₁₈ column is a suitable and practical method for the isolation of alkaloids in plants.     

A LC‐MS/MS method for simultaneous determination of seven alkaloids in rat plasma after oral administration of Phellodendri chinensis cortex extract and its application to a pharmacokinetic study

A sensitive and rapid liquid chromatography with tandem mass spectrometry method has been developed for simultaneous determination of berberine (I), jateorhizine (II), palmatine (III), tetrahydropalmatine (IV), phellodendrine (V), protopine (VI) and columbamine (VII) in rat plasma after oral administration of Phellodendri chinensis cortex extraction. The plasmas were extracted by liquid‐liquid extraction. The tandem mass spectrometric detection was performed in the multiple reaction monitoring mode in the positive ionization. The intra‐ and interday precisions and accuracies were in range from ?12.18 to 13.21%. Mean absolute recoveries of all analytes and internal standard were between 78.6 and 98.9%. The seven alkaloids were proven to be stable during sample storage and analysis procedures. The established method was validated and successfully applied to pharmacokinetics study in rat plasma after oral administration of Phellodendri chinensis cortex extract. The t_1/2 of palmatine, columbamine, pellodendrine, berberine, tetrahydropalmaine, jatrorrhizine, and protopine were 5.16, 5.96, 7.18, 19.84, 6.28, 7.08, 6.90 h, respectively. The seven compounds could be rapidly absorbed into blood (time for maximal concentration, 1.80–1.93 h). This study could establish a foundation for further research of Phellodendri chinensis cortex and might provide more useful information to guide the clinical usage.     

Evaluation of alkaloids binding to the parallel quadruplex structure [d(TGGGGT)]4 by electrospray ionization mass spectrometry

In this study, electrospray ionization mass spectrometry (ESI‐MS) was used to investigate the binding interaction of six alkaloids with parallel intermolecular G‐quadruplex [d(TGGGGT)]₄, and five alkaloids including berberine, jatrorrhizine, palmatine, tetrandrine, and fangchinoline showed complexation with the target DNA. Relative binding affinities were estimated on the basis of mass spectrometric data. The slight differences in chemical structures of berberine, jatrorrhizine, and palmatine had little influence on their binding affinities to [d(TGGGGT)]₄. Tetrandrine and fangchinoline selectively bound to [d(TGGGGT)]₄ versus duplex DNA. Collision‐induced dissociation (CID) experiments showed that the complexes with berberine, jatrorrhizine, and palmatine dissociated via strand separation and ligand retaining in the strand while the complexes with tetrandrine and fangchinoline were dissociated via ligand elimination. A comparison of dissociation patterns in CID experiments of complexes with the alkaloids to those with the traditional G‐quadruplex DNA binders suggested an end‐stacking binding mode for tetrandrine and fangchinoline and an intercalation binding mode for berberine, jatrorrhizine, and palmatine to the target DNA. The current work not only provides deep insight into alkaloid/[d(TGGGGT)]₄ complexes and useful guidelines for design of efficient anticancer agents but also demonstrates the utility of ESI‐MS as a powerful tool for evaluating interaction between ligand and quadruplex DNA. Copyright © 2012 John Wiley & Sons, Ltd.     

Simultaneous determination of eight bioactive components in rat plasma after oral administration of Yan‐Ke‐Ning‐Tablet by liquid chromatography coupled with Q‐Exactive Orbitrap tandem mass spectrometry

A rapid, sensitive and reliable quantitative method based on ultra‐high performance liquid chromatography coupled with Q‐Exactive Orbitrap tandem mass spectrometry was developed for simultaneous determination of berberine, berberrubine, palmatine, jatrorrhizine, columbamine, baicalin, baicalein and wogonin in rat plasma after oral administration with Yan‐Ke‐Ning‐Tablet (YKNT). After precipitation with acetonitrile, the plasma samples were separated on a reverse‐phase C₁₈ column with 1 mm ammonium acetate containing 0.2% acetic acid–acetonitrile as mobile phase. Calibration curves showed good linearity (r > 0.9983) over the tested concentration ranges of 0.5–200 ng/mL for berberine, berberrubine, palmatine, jatrorrhizine and columbamine, and 1–300 ng/mL for baicalin, baicalein and wogonin. The precision (relative standard deviation) at three different concentration levels was <12.15% and the accuracy (relative error) ranged from ?8.24 to 10.85%. No matrix effects were observed with matrix effect value ranging from 89.23 to 107.68%. The extraction recovery was in the range of 82.34–92.31%. The validated assay was further successfully applied to the pharmacokinetic study of these components after oral administration of YKNT. The present study provides the pharmacokinetic profiles of major bioactive components found in YKNT, and provides valuable information regarding the chemical components that were absorbed into plasma, which will be helpful for understanding the therapeutic effects of YKNT.     

Echinophora tenuifolia L. branches phytochemical profile and antiproliferative activity on human cancer cell lines

Abstract

The methanolic extract of Echinophora tenuifolia L. branches and its fractions were evaluated for their in vitro cell growth inhibitory activity on different human cancer cell lines (C32, LoVo and SKBr3) and the normal BJ fibroblasts. All tested samples were effective against the melanoma cell line C32, with IC₅₀ values ranging from 22.8?±?0.8 to 78.7?±?1.2?μg/mL, the antiproliferative activity of the dichloromethane fraction being significantly higher. This fraction was also effective against the LoVo adenocarcinoma cell line, with an IC₅₀ value of 53.0?±?2.1?μg/mL. The ethyl acetate and dichloromethane fractions showed the highest lipid peroxidation inhibitory activity, verified by means of the β-carotene bleaching test. The phytochemical profiles of E. tenuifolia branches extract were established by means of GC-MS and HPTLC. Overall, branches of E. tenuifolia L. could represent a rich source of bioactive compounds, potentially useful in the pharmaceutical field.     

Novel molecularly imprinted magnetic nanoparticles for the selective extraction of protoberberine alkaloids in herbs and rat plasma

In this work, a novel magnetic nanomaterial functionalized with a molecularly imprinted polymer was prepared for the extraction of protoberberine alkaloids. Molecularly imprinted polymers were made on the surface of Fe₃O₄ nanoparticles by using berberine as template, acetonitrile/water as porogen, acrylamide as functional monomer and ethylene glycol dimethacrylate as cross‐linker. The optimized molar ratio of template/functional monomer was 1:7. The polymeric magnetic nanoparticles were characterized by transmission electron microscopy and Fourier transform infrared spectroscopy. The stability and adsorption capacity of the molecularly imprinted polymers were investigated. The molecularly imprinted polymers were used as a selective sorbent for the magnetic molecularly imprinted solid‐phase extraction and determination of jatrorrhizine, palmatine, and berberine. Extraction parameters were studied including loading pH, sample volume, stirring speed, and extraction time. Finally, a magnetic molecularly imprinted solid‐phase extraction coupled to high‐performance liquid chromatography method was developed. Under the optimized conditions, the method showed good linear range of 0.1–150 ng/mL for berberine and 0.1–100 ng/mL for jatrorrhizine and palmatine. The limit of detection was 0.01 ng/mL for berberine and 0.02 ng/mL for jatrorrhizine and palmatine. The proposed method has been applied to determine protoberberine alkaloids in Cortex phellodendri and rat plasma samples. The recoveries ranged from 87.33–102.43%, with relative standard deviation less than 4.54% in Cortex phellodendri and from 102.22–111.15% with relative standard deviation less than 4.59% in plasma.     

Antileishmanial diterpenoid alkaloids from Aconitum spicatum (Bruhl) Stapf

The crude extracts of tubers of Aconitum spicatum (Bruhl) Stapf were investigated for in vitro antileishmanial activity against Leishmania major. The dichloromethane extract at pH 2.5 showed antileishmanial activity with IC₅₀ value of 27.10 ± 0.0 μg/mL. Chromatographic purification of the dichloromethane extract led to isolation of three C-19 norditerpenoid alkaloids indaconitine (1), chasmaconitine (2) and ludaconitine (3). Compounds 3 and 2 showed antileishmanial activity with IC₅₀ = 36.10 ± 3.4 and 56.30 ± 2.1 μg/mL, respectively. Compound 1 was less effective (IC₅₀ > 100 μg/mL). The cytotoxicity of compounds 1, 2 and 3 studied against MCF7, HeLa and PC3 cancer cell lines and 3T3 normal fibroblast cell line did not show cytotoxicity at 30 μM.     

α-Glucosidase Inhibition and Antioxidant Properties of Streptomyces sp.: In Vitro

Streptomyces strain isolated from the soil sediment was studied for its in vitro α-glucosidase and antioxidant properties. Morphological characterization and 16S rRNA partial gene sequencing were carried out to confirm that the strain Loyola AR1 belongs to genus Streptomyces sp. Modified nutrient glucose broth was used as the basal medium for growth and metabolites production. Ethyl acetate extract of Loyola AR1 (EA-Loyola AR1) showed 50 % α-glucosidase inhibition at the concentration of 860.50?±?2.68 μg/ml. Antioxidant properties such as total phenolic content of EA-Loyola AR1 was 176.83?±?1.17 mg of catechol equivalents/g extracts. EA-Loyola AR1 showed significant scavenging activity on 2,2-diphenyl-picrylhydrazyl (50 % inhibition (IC₅₀), 750.50?±?1.61 μg/ml), hydroxyl (IC₅₀, 690.20?±?2.38 μg/ml), nitric oxide (IC₅₀, 850.50?±?1.77 μg/ml), and superoxide (IC₅₀, 880.08?±?1.80 μg/ml) radicals, as well as reducing power. EA-Loyola AR1 showed strong suppressive effect on lipid peroxidation (IC₅₀, 670.50?±?2.52 μg/ml). Antioxidants of β-carotene linoleate model system reveals significantly lower than butylated hydroxyanisole.     

Evaluation of the cytotoxicity on breast cancer cell of extracts and compounds isolated from Hyptis pectinata (L.) poit

Abstract

Hyptis pectinata is a herb popularly used in Brazil for the treatment of inflammations, pain, bacterial infections and cancer. In the present study, inflorescences (MPIn), leaves (MPL), branches (MPB), root (MPR) extracts and three compounds isolated from MPIn were assayed against breast tumor cell lines. The structures of the three compounds (pectinolide J, hyptolide and pectinolide E) were determined by means of spectroscopic analysis. Pectinolide J was isolated for the first time. The MPIn, MPL and MPR exhibited specific antiproliferative activity on tumor cell lines when compared to normal cell lines with IC₅₀ of 52.01?±?0.64, 45.91?±?0.02?μg/mL and 82.84?±?0.03?μg/mL, respectively. Although the isolated substances did not present good antiproliferative activity, when the three were associated, a greater biological effect was observed, suggesting a synergistic effect. Hyptolide (5.6?±?0.4?μg/mL) showed IC₅₀ sufficiently low to be considered as a drug prototype.     

Investigation of the effect of berberines alkaloids in <Emphasis Type="Italic">Coptis chinensis</Emphasis> Franch on <Emphasis Type="Italic">Bacillus shigae</Emphasis> growthby microcalorimetry

The inhibitory effects of five berberines alkaloids (BAs) from rhizoma of Coptis chinensis Franch, a traditional Chinese medicinal (TCM) herb, on Bacillus shigae (B. shigae) growth were investigated by microcalorimetry. The power-time curves of B. shigae with and without BAs were acquired; meanwhile, the extent and duration of inhibitory effects on the metabolism were evaluated by growth rate constants (k1, k2), half inhibitory ratio (IC50), maximum heat output (Pmax), and peak time (tp). The values of k1 and k2 of B. shigae in the presence of the five BAs decreased with the increasing concentrations of BAs. Moreover, Pmax was reduced, and the value of tp increased with increasing concentrations of the five drugs. The inhibitory activity varied with different drugs. IC50 of the five BAs was respectively 75 μg/mL for berberine, 90 μg/mL for coptisine, 115 μg/mL for palmatine, 220 μg/mL for epiberberine, and 400 μg/mL for jatrorrhizine. The sequence of antimicrobial activity of the five BAs berberine ＞ coptisine ＞ palmatine ＞ epiberberine ＞ jatrorrhizine. The functional groups methylenedioxy at C2 and C3 on phenyl ring improve antimicrobial activity more strongly than methoxyl at C2 and C3 on phenyl ring. However, the effect of bacteriostasis is not significant with methylenedioxy or methoxyl at C9 and C10 on phenyl ring.     

Antioxidant,cholinesterase inhibition activities and essential oil analysis of Nelumbo nucifera seeds

Nelumbo nucifera seeds’ essential oil (EO), crude extract and subsequent fractions were evaluated for their DPPH, ABTS and superoxide anion-free radical scavenging and cholinesterase inhibitory activities. The ethyl acetate fraction and EO showed outstanding antioxidant activities with IC₅₀ values of 191, 450 μg/mL (DPPH), 123, 221 μg/mL (ABTS) and 69, 370 μg/mL (superoxide anion). The ethyl acetate fraction and EO also caused significant inhibition of acetylcholinesterase and butyrylcholinesterase with IC₅₀ values of 70 ± 0.6, 64 ± 0.8 and 75 ± 0.3, 58 ± 0.2, in dose-dependent manner. The first ever gas chromatography–mass spectrometry analysis of the EO obtained from N. nucifera seeds resulted in identification of 19 constituents, mainly comprised of oxygenated sesquiterpenes responsible for their promising bioactivity. The crude and fractions revealed the presence of saponins, flavonoids, steroids, alkaloids, terpenoids and cardiac glycosides in phytochemical investigation.     

Comparative study of the phenolic profile,antioxidant and antimicrobial activities of leaf extracts of five Juniperus L. (Cupressaceae) taxa growing in Turkey

Abstract

The purpose of this study was to conduct a comparative evaluation of some biological properties of methanol and water extracts of leaves of five Juniperus taxa growing in Turkey: J. communis L. var. communis (Jcc), J. communis L. var. saxatilis Pall. (Jcs), J. drupacea Labill. (Jd), J. oxycedrus L. subsp. oxycedrus (Joo), J. oxycedrus L. subsp. macrocarpa (Sibth. & Sm.) Ball. (Jom). The antioxidant properties were examined in vitro; both in the DPPH and in the reducing power tests, Joo methanol extract resulted the most active (IC₅₀?=?0.09?±?0.01mg/mL and ASE/mL?=?2.56?±?0.06). In the TBA assay, Jcs methanol extract exhibited the highest activity (IC₅₀?=?4.39?±?0.47?μg/mL). The extracts displayed bacteriostatic activity against Staphylococcus aureus, and Jd methanol extract resulted the most effective (MIC?=?19.53?μg/mL); no effect on the S. aureus biofilm formation was observed. The extracts resulted non-toxic in the Artemia salina lethality bioassay. Finally, the phenolic profile of the methanol extracts was characterized by HPLC-PDA/ESI-MS.     

Chemical constituents of essential oil of endemic Rhanterium suaveolens Desf. growing in Algerian Sahara with antibiofilm,antioxidant and anticholinesterase activities

Twenty compounds were detected in the essential oil of Rhanterium suaveolens representing 98.01% of the total oil content. Perillaldehyde (45.79%), caryophyllene oxide (24.82%) and β-cadinol (5.61%) were identified as the main constituents. In β-carotene–linoleic acid assay, both the oil and the methanol extract exhibited good lipid peroxidation inhibition activity, with IC₅₀ values of 17.97 ± 5.40 and 11.55 ± 3.39 μg/mL, respectively. In DPPH and CUPRAC assays, however, the methanol extract exhibited a good antioxidant activity. The highest antibiofilm activity has been found 50.30% against Staphylococcus epidermidis (MU 30) at 20 μg/mL for essential oil and 58.34% against Micrococcus luteus (NRRL B-4375) at 25 mg/mL concentration for methanol extract. The in vitro anticholinesterase activity of methanol extract showed a moderate acetylcholinesterase inhibitory (IC₅₀ = 168.76 ± 0.62 μg/mL) and good butyrylcholinesterase inhibitory (IC₅₀ = 54.79 ± 1.89 μg/mL) activities. The essential oil was inactive against both enzymes.     

Inula viscosa (L.) Aiton leaves and flower buds: Effect of extraction solvent/technique on their antioxidant ability,antimicrobial properties and phenolic profile

Abstract

This study was designed to establish the most effective solvent/technique for extracting antioxidant phytoconstituents from leaves and flower buds of Inula viscosa (L.) Aiton (Asteraceae) grown wild in Morocco. Maceration and hot extraction with methanol or water and Soxhlet ethanol extraction were utilized. The antioxidant potential was evaluated in vitro by DPPH, reducing power, and ferrous ions chelating activity assays. I. viscosa leaf and flower bud extracts displayed the strongest effect in the DPPH test, being the Soxhlet ethanol the most active ones (IC₅₀ = 54.24?±?0.21?μg/mL and 39.77?±?0.23?μg/mL); thus, they were selected for further investigations. The antimicrobial efficacy of the Soxhlet ethanol extracts against ATCC and food isolates strains was assayed; the leaf extract showed the best activity, and Candida albicans was the most sensitive strain (MIC = 125?µg/mL). The extracts resulted non-toxic against Artemia salina. Among the phenolics characterised by HPLC-PDA-ESI-MS, hispidulin hexoside, patuletin and spinacetin were identified for the first time.     

Synthesis,characterization, and hypoglycemic efficacy of nitro and amino acridines and 4-phenylquinoline on starch hydrolyzing compounds: an in silico and in vitro study

α-Amylase and α-Glucosidase are important therapeutic targets for type II diabetes. The present focus of our study is to elucidate the hypoglycemic activity of novel compounds through in vitro and in silico studies. Here, we synthesized the nitro acridines (3a–3c), amino acridines (4a–4c), and nitro phenylquinoline (3d) and amino phenylquinoline (4d) using a multi-step reaction protocol in good yields. All the above derivatives were screened for molecular docking, α-Amylase and α-Glucosidase inhibitory activities utilizing acarbose as standard drug. In silico studies were performed to explore the binding ability of compounds with the active site of α-Amylase and α-Glucosidase enzymes. The in vitro antihyperglycemic report of 3c exhibits the maximum inhibitory activity with IC₅₀ values of 200.61?±?9.71 μmol/mL and 197.76?±?8.22 μmol/mL against α-Amylase and α-Glucosidase, respectively. Similarly, the compound 3a exhibits IC₅₀ values of 243.78?±?13.25 μmol/mL and 296.57?±?10.66 μmol/mL, and 4c exhibits IC₅₀ values of 304.28?±?3.51 μmol/mL and 278.86?±?3.24 μmol/mL with a significant p?<?0.05 in both enzyme inhibitions. In addition, the presence of diverse functional moieties in synthesized compounds may provide a strong inhibitory action against the abovementioned enzymes compared with standard acarbose inhibition (IC₅₀, 58.74?±?3.68 μmol/mL and 49.39?±?4.94 μmol/mL). Also, the docking studies provided an excellent support for our in vitro studies. The outcome of these studies recommends that the tested compounds might be treated as potential inhibitors for the starch hydrolyzing enzymes in type II diabetes.

     

Determination of Protoberberine Alkaloids in Coptis chinensis by Microextraction and High Performance Liquid Chromatography

Sample preparation technique based on an organic filter membrane (pH-resolved filter membrane microextraction) (pH-RFMME) was developed, coupled with high-performance liquid chromatography, and used to determine protoberberine alkaloids (jatrorrhizine, epiberberine, coptisine, palmatine, and berberine) in Coptis chinensis at different pH values through a one-step procedure. This green procedure provides a desirable sample pretreatment technology. The main variables affecting the extraction such as filter membrane area (or volumes of extraction solvents), sample pH, eluent pH, ionic strength, extraction stirring rate, extraction time, and sample volume were optimized. Under the optimized conditions, the enrichment factors of the analytes were 40.4–52.0, the linear ranges were 3.2–6250 ng · mL^?1 for jatrorrhizine and epiberberine, 6.0–12000 ng · mL^?1 for coptisine, 1.8–3600 ng · mL^?1 for palmatine, and 18.8–18800 ng · mL^?1 for berberine, with r ² ≥ 0.9945. The limits of detection were less than 0.3 ng · mL^?1. Satisfactory recoveries (84.8%–115.5%) and precision (1.8%–10.0%) were also achieved. These results confirmed that pH-RFMME is a simple, rapid, practical, and environmentally friendly method to isolate analytes that exhibit significant differences in acidity or alkalinity from complex samples.     

Antioxidant,antimicrobial and urease inhibiting activities of methanolic extracts from Cyphostemma digitatum stem and roots

Cyphostemma digitatum stem and roots extracts were investigated for antioxidant, antimicrobial, urease inhibition potential and phytochemical analysis. Phytochemical screening of the roots and stem extract revealed the presence of secondary metabolites including flavonoids, alkaloids, coumarins, saponins, terpenoids, tannins, carbohydrates/reducing sugars and phenolic compounds. The methanolic extracts of the roots displayed highest antioxidant activity (93.518%) against DPPH while the crude methanolic extract of the stem showed highest antioxidant activity (66.163%) at 100 μg/mL concentration. The methanolic extracts of both stem and roots were moderately active or even found to be less active against the selected bacterial and fungal strains (Tables S2 and S3). The roots extract (methanol) showed significant urease enzyme inhibition activity (IC₅₀ = 41.2 ± 0.66; 0.2 mg/mL) while the stem extract was found moderately active (IC₅₀ = 401.1 ± 0.58; 0.2 mg/mL) against thiourea (IC₅₀ = 21.011; 0.2 mg/mL).